James p allison, 1996 enhancement of antitumor immunity by CTLA 4

American Association for the Advancement of Science http://www.jstor.org/stable/2890840 Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at http://www.jstor.org/page/info/about/policies/terms.jsp JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive We use information technology and tools to increase productivity and facilitate new forms of scholarship For more information about JSTOR, please contact support@jstor.org American Association for the Advancement of Science is collaborating with JSTOR to digitize, preserve and extend access to Science http://www.jstor.org This content downloaded from 128.32.19.34 on Wed, 17 Jul 2013 12:44:45 PM All use subject to JSTOR Terms and Conditions try and 100% of whom were of other ethnic backgrounds The stop codon mutation was screened in 70 Finnish EPM1 carrier parents All 70 of these individualscontained the common ancestral haplotype around the EPM1 locus on one of their chromosomes To distinguish mutations from polymorphisms, we considered only the nonancestral haplotype chromosome of these 70 individuals.DNAfrom these individuals was amplified by PCR, and the products were directly sequenced with the AmpliCycle sequencing kit (Perkin-Elmer) 17 R Jerala, M Trstenjak, B Lenarcic, V Turk, FEBS Lett 239, 41 (1988) 18 M Abrahamson, M Q Islam, J Szpirer, C Szpirer, G Levan, Hum Genet 82, 223 (1989); J Ghiso, Jensson, B Frangione,Proc Natl Acad Sci U.S.A 83, 2974 (1986) 19 R Eldridge,M livanainen,R Stern, T Koerber, B J Wilder,Lancet ii, 838 (1983) 20 We thank the families with EPM1 for contributingto this study; C lannicola,C Prange, D Vollrath,J Kere, and members of the Myers and Cox laboratoriesand the Stanford Human Genome Center for discussions and support; A.-L.Traskelinand R Tolvanenfortechnical assistance; and R Eldridgeand B J Wilderfor providingpatient samples from the American family This work was supported by NIH grants HD-24610 and P50 HG-00206 (to R.M.M.and D.R.C.),postdoctoral grant NIH IF32GM17502 (to J.A.W.), NIHgrant NS31831 (to A.d.I.C.),the Academy of Finlandand the Sigrid Juselius Foundation (to A.d.1.C and A.E.L.), and the Epilepsy Research Foundation of Finland (to A.-E.L.) Part of this study was done at the FolkhalsanInstituteof Genetics (Helsinki) 26 January 1996; accepted 14 February1996 Enhancement of Antitumor Immunity by CTLA-4Blockade Dana R Leach, Matthew F Krummel, James P Allison* One reason for the poor immunogenicity of many tumors may be that they cannot provide signals for CD28-mediated costimulation necessary to fully activate T cells It has recently become apparent that CTLA-4, a second counterreceptor for the B7 family of costimulatory molecules, is a negative regulator of T cell activation Here, in vivo administration of antibodies to CTLA-4 resulted in the rejection of tumors, including preestablished tumors Furthermore,this rejection resulted in immunity to a secondary exposure to tumor cells These results suggest that blockade of the inhibitory effects of CTLA-4 can allow for, and potentiate, effective immune responses against tumor cells 20E Fig 15 - Control Days after tumor-iAnti-CD28 wt anti-CTLA-4 Anti-CTLA-4 reten t t N - 10E 05 06 10 15 2'0 2'5 3'0 35 60 Days aftertumor injection Fig Treatmentwith anti-CTLA-4accelerates rejection of a B7-1 -positive colon carcinoma (23) A volume of 100 lI of cell suspension (4 x 106 cells) was injected subcutaneously into the left flanks of groups of five female BALB/c mice Two of the groups received three intraperitonealinjections of either anti-CTLA-4 or anti-CD28 (18) Injections of 100, 50, and 50 pLgof antibody were given on days 0, 3, and 6, respectively, as indicated by the arrows Control animals received no injections Data points represent the average of the products of bisecting tumor diameters Error bars represent standard errorof the mean bivaCTLA-4 blockade with nonstimnulatory, lent antibody (18, 20) would accelerate rejection of B7-positive tumor cells Previously, we showed that B7-1 expression was partially successful at inducing rejection of the transplantable murine colon carcinoma 51 BLimlI0 (23) We reasoned that CTLA-4 blockade Despite expressingantigens recognizable tumorcells transfectedwith B7 are able to would remove inhibitory signals in the coby a host'simmunesystem,tumorsarevery behave as APCs, presumablyallowing di- stimulatory pathway, resulting in enhanced poorin initiatingeffectiveimmunerespons- rect activationof tumor-specificT cells rejection of the tumor cells We injected es One reasonfor this poor immunogenicRecent evidence suggeststhat costimu- groups of BALB/c mice with B7-1-transity maybe that the presentationof antigen lation is more complex than originally fected lBLimlO tumorcells (B7-5 1BLiml0) alone is insufficientto activate T cells In thoughtand involvescompetingstimulato- (23) Two groupswere treated with a series of intraperitoneal injections of either antiadditionto T cell receptorengagementof ry and inhibitory signaling events (3, an antigenicpeptideboundto majorhisto- 9-12) CTLA-4,a homologof CD28, binds CTLA-4 or anti-CD28 (18, 24) Treatment compatibilitycomplex (MHC) molecules, both B7-1 and B7-2 with affinities much with anti-CTLA-4 inhibited B7-51BLimlO additionalcostimulatorysignals are neces- greaterthan does CD28 (13-16) In vitro, tumor growth as compared with the antisary for T cell activation (1) The most antibodycross-linkingof CTLA-4has been CD28-treated mice or the untreatedcontrols importantof these costimulatorysignalsap- shown to inhibit T cell proliferationand (Fig 1) All mice in the untreated and antipears to be providedby the interactionof interleukin-2productioninduced by anti- CD28-treated groupsdeveloped small tumors CD28 on T cells with its primaryligands bodyto CD3 (anti-CD3),whereasblockade that grew progressivelyfor to 10 days and B7-1 (CD80) and B7-2 (CD86) on the of CTLA-4 with solubleintact or Fabfrag- then ultimately regressedin of the 10 mice surface of specialized antigen-presenting ments of antibody enhances proliferative by about day 23 after injection The two small cells (APCs) (2-4) Expressionof B7 coresponses(17, 18) Similarly,solubleintact tumorsthat did not regressremainedstatic for or Fab fragmentsof anti-CTLA-4 greatly more than 90 days In contrast, tlhreeof five stimulatory molecules is limited to specialized APCs Therefore,even though most augmentT cell responsesto nominal pep- mice treated witlh anti-CTLA-4 developed tissue-derivedtumorsmay present antigen tide antigen or the superantigenStaphylo- very small tumors,all of which regressedcomin the context of MHCmolecules,they may coccusenterotoxinB in vivo (19, 20) It has pletely by day 17 Although these resultswere fail to elicit effectiveimmunitybecauseof a also been suggestedthat CTLA-4 engage- encouraging and were consistent with our lack of costimulatory ability Several studies hypothesis, tlley were not very dramatic bement can induce apoptosisin activatedT cause B7-1 expression resulted in fairly rapid supportthis notion In a varietyof model cells (21) Finally, mice deficient in systems,transfectedtumorcells expressing CTLA-4 exhibit severeT cell proliferative rejection of transfected 51BLimlO cells even costimulatoryB7 moleculesinducedpotent disorders(22) These results demonstrate in the absence of CTLA-4 blockade; however, these resultsconfirmed that anti-CTLA-4 responsesagainst both modified and un- that CTLA-4 is a negative regulatorof T modifiedtuLmor cells (5-8) It appearsthat cell responsesand raise the possibilitythat did not inhibit tumor rejection blockadeof inhibitorysignalsdeliveredby WXe next examined the effects of CTLA-4 blockade on the growth of T augment interactions might CTLA-4-B7 Cancer Research Laboratoryand Department of Moleccell responsesto tumor cells and enhance V51BLiml1O, a vector control tumor cell ular and Cell Biology, University of California,Berkeley, CA 94720, USA line that does not express B7 (23) All mice antitumor immunity *To whom correspondence should be addressed We first sought to determinewhether either injected with x 10o6 V51BLiml1O 1734 SCIENCE * VOL 271 * 22 MARCH 1996 This content downloaded from 128.32.19.34 on Wed, 17 Jul 2013 12:44:45 PM All use subject to JSTOR Terms and Conditions R:PORTS 200 A - 175 E 150 -0 150 BC j Control Anti-CD28 Anti-CTLA-4 T3 40 25 125 (I, Anti-CTLA-4 -e_ Control Anti-CTLA-4 -p E 125- I 75 ,v ,t I 10 15 20 25 30 35 40 90 Days aftertumor injection t 50100000000123456789 t (D~~~~~~~~~~~~~~~~~~~~7 Days after tumor injection 200 D Fig Treatment with anti-CTLA-4 enhances rejection of B7-negative colon carcinoma ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ _- V5IBLIm*Oe 50cells and results in protection against subsequent challenge with wild-type colon carciAnti-CTLA-4 e 15 noma cells Groups of BALB/c mice were injected with B7-negative 51 BLimi10vector E 150 -e- ChallengeonlyX control cells (V51BLim10), left untreated, or treated with anti-OTLA-4or control antibody ulcerated If 125 Mice were euthanized when tumors reached a size of 200 mm2 or became en individualmice withina group were euthanized, the finalmeasurement was carriedover to 100 l subsequent time points (A)Average tumor size in mice injected with x 1Q6 tumor cells s 750 tumorcells Treatedgroups Groupsof fivemicewere injectedwith4 x 1Q6 V51BLim1 or anti-0D28as indicatedby the were injectedthreetimeswith100 pFgof anti-OTLA-4 0>5